节旋藻(螺旋藻)高分子量DNA的两种制备方法  

TWO PROCOTOLS FOR PREPARATION OF HIGH MOLE- CULAR WEIGHT DNA FROM Arthrospira

作　　者：茅云翔[1] 张宝红[1] 杨官品[1] 张学成[1] 

机构地区：[1]青岛海洋大学海洋生命学院,青岛266003

出　　处：《海洋科学》2003年第2期32-36,共5页Marine Sciences

基　　金：国家863计划资助项目 (819 06- 10)

摘　　要：DNA的简便高效制备方法是研究基因结构、功能及开展其它各项研究的基础。首次报道了针对节旋藻的结构和组成特性而建立的制备节旋藻高分子量DNA的两种方法。其中第一个方法是常规方法 ,可大量提取纯度较高、分子量达50kb的节旋藻DNA ,可用于构建节旋藻质粒文库、Southern杂交和PCR操作等 ;第2种方法是用脉冲电场凝胶电泳分离DNA片段 ,制备的DNA片段达数百kb ,可用于构建节旋藻噬菌体文库、粘粒文库、BAC文库等 ,从而为构建节旋藻物理图谱 ,定位克隆基因奠定基础。Simple and high efficiency methods for DNA preparation are the base for the molecular genetics research. Here, for the first time, two protocols were designed for preparing the high weight molecular DNA from Arthrospira based on its structural and biochemical characters. The first protocol can be used for routine and large-scale DNA preparation, by which the molecular weight of DNA is more than 50 kb. The DNA of Arthrospira prepared by this protocol can be used for constructing plasmid library, Southern blotting and PCR experimenting. By the second protocol, large DNA fragments over several hundred of kilobasepairs can be obtained by the pulse field gel electrophoresis, which can be used for constructing ( phage library, cosmid library and bacterial artificial chromosome (BAC) library, and thereby for constructing physical map and positional clone gene of Arthrospira.

关 键 词：节旋藻 螺旋藻 高分子量DNA 制备方法 脉冲场凝胶电泳 

分 类 号：Q949.221[生物学—植物学] Q943